The structures of twelve ester-type polyoxypregnane derivatives, isolated from Marsdenia tomentosa DECNE (Asclepiadaceae), were determined with some interesting information for their properties on the bases of chemical and physico-chemical evidences. The configuration at C-20 of tomentogenin (I) and utendin (II) was determined as S by means of the circular dichroism (CD) experiment of 20-O-O-nitrobenzoyl-derivatives to define their absolute configurations. A hypothetical biogenetic pathway of polyoxypregnanes in M. tomentosa is proposed on the bases of the position of ester linkages and the nature of oxygen functions of ester-type derivatives.